Ander Pino

Biological Stability of Plasma Rich in Growth Factors Eye Drops After Storage of 3 Months.

Purpose: We evaluated whether plasma rich in growth factors eye drops maintain their composition and biological activity after storage for 3 months at −20°C and after storage at 4°C or room temperature (RT) for 24 hours, compared with samples obtained at time 0 (fresh samples). Methods: Blood from 10 healthy donors was collected, centrifuged, and plasma rich in growth factors was prepared by avoiding the collection of the buffy coat. Eye drops were kept fresh or were stored at −20°C for 15, 30, and 90 days. For each time, 2 aliquots were kept at RT or at 4°C for 24 hours. Osmolarity, vitamin A, fibronectin, platelet-derived growth factor-AB, vascular endothelial growth factor, epithelial growth factor, and transforming growth factor-&bgr;1 were quantified. The proliferative and migratory potential of the eye drops was assayed on primary human keratocytes. Results: Platelet-derived growth factor-AB, vascular endothelial growth factor, epithelial growth factor, and vitamin A levels remained constant for each time and for each storage condition, whereas fibronectin, transforming growth factor-&bgr;1, and osmolarity values were slightly modified after freezing. Cell proliferation and migration were significantly enhanced with the biological eye drops independently of the time and the storage condition. No microbial contamination was observed in any plasma rich in growth factors eye drops. Conclusions: Plasma rich in growth factors eye drops can be stored for up to 3 months without any reduction of the main proteins involved in ocular surface healing. Their use during 24 hours either at 4°C or at RT did not alter the composition and the in vitro biological activity of the eye drops.

The Effect of Plasma Rich in Growth Factors on Pattern Hair Loss: A Pilot Study

BACKGROUND Autologous growth factors have proved to promote tissue regeneration in various medical fields. Recent findings suggest that platelet rich plasma may also play an important role in hair follicle restoration. OBJECTIVE The objective of this study was to evaluate the safety and clinical efficacy of plasma rich in growth factors (PRGFs) for the treatment of androgenetic alopecia (AGA). MATERIALS AND METHODS Five PRGF injections were administered over 19 patients with AGA. Phototrichograms regarding follicle density/diameter and terminal/vellus hair ratio were performed at baseline and after 1 year follow-up period. Consenting participants underwent histologic scalp examination. At the end of the study, overall patient satisfaction and clinical improvement were determined. RESULTS After PRGF therapy, mean hair density/diameter increased and terminal/vellus hair ratio was also improved. Patients presented epidermal thickness, perifollicular neoangiogenesis, cell proliferation, and terminal/miniaturized hair ratio improvement. Plasma rich in growth factors seemed to reduce the perivascular inflammatory infiltrate, promote the remodeling of dermo-epidermal tissue, and increase bulge stem cell niches. Patients declared an overall positive satisfaction, and a high clinical improvement score was achieved when comparing premacrophotographs and postmacrophotographs. CONCLUSION Although randomized clinical trials are needed, this study provides preliminary data supporting the positive therapeutic effect of autologous growth factors on hair follicle regeneration.

Implementation of a more physiological plasma rich in growth factor (PRGF) protocol: Anticoagulant removal and reduction in activator concentration

Abstract Plasma rich in growth factors (PRGF) is a biological therapy that uses patient’s own growth factors for promoting tissue regeneration. Given the current European regulatory framework in which anticoagulant solution in blood extraction tubes could be considered as a medicinal product, a new PRGF protocol has been developed. The actual protocol (PRGF-A) and the new one (PRGF-B) have been performed and compared under Good Laboratory Practices. PRGF-A protocol uses extraction tubes with 0.9 mL of trisodium citrate as anticoagulant and 50 μL of calcium chloride/mL PRGF to activate it. The PRGF-B reduces the amount of sodium citrate and calcium chloride to 0.4 mL and to 20 μL, respectively. Basic hematological parameters, platelet function, the scaffold obtaining process, growth factors content, and the biological effect were compared between both PRGF obtaining protocols. Results: PRGF-B protocol led to a statistically significant higher enrichment and recovery of platelets regarding to the PRGF-A. Hypotonic stress response by platelets was significantly better in the new protocol. A statistically significant decrease in the basal platelet activation status of PRGF-B compared to PRGF-A was also observed. The duration of the lag phase in the platelet aggregation assay was statistically lower for the PRGF-B protocol. Both the clotting and the clot retraction time were significantly reduced in the B protocol. A higher growth factor concentration was detected in the plasma obtained using the PRGF-B protocol. The new PRGF obtaining protocol, with a reduction in the amount of anticoagulant and activator, has even improved the actual one.

Plasma Rich in Growth Factors Enhances Wound Healing and Protects from Photo-oxidative Stress in Dermal Fibroblasts and 3D Skin Models.

BACKGROUND Optimal skin repair has been a desired goal for many researchers. Recently, plasma rich in growth factors (PRGF) has gained importance in dermatology proving it is beneficial effects in wound healing and cutaneous regeneration. OBJECTIVE The anti-fibrotic, pro-contractile and photo-protective effect of PRGF on dermal fibroblasts and 3D skin models has been evaluated. METHOD The effect against TGFβ1 induced myofibroblast differentiation was tested. Cell contractile activity over collagen gel matrices was analyzed and the effect against UV derived photo-oxidative stress was assessed. The effectiveness of PRGF obtained from young aged and middle aged donors was compared. Furthermore, 3D organotypic skin explants were used as human skin models with the aim of analyzing ex vivo cutaneous preventive and regenerative photo-protection after UV exposure. RESULTS TGFβ1 induced myofibroblast levels decreased significantly after treatment with PRGF while the contractile activity increased compared to the control group. After UV irradiation, cell survival was promoted while apoptotic and ROS levels were noticeably reduced. Photo-exposed 3D explants showed higher levels of metabolic activity and lower levels of necrosis, cell damage, irritation and ROS formation when treated with PRGF. The histological integrity and connective tissue fibers showed lower signals of photodamage among PRGF injected skin models. No significant differences for the assessed biological outcomes were observed when PRGF obtained from young aged and middle aged donors were compared. CONCLUSION These findings suggest that this autologous approach might be useful for antifibrotic wound healing and provide an effective protection against sun derived photo-oxidative stress regardless the age of the patient.

Opening new horizons in regenerative dermatology using platelet-based autologous therapies

Biological therapeutic therapies are gaining the attention of scientists and medical doctors. Accumulating evidence suggests that blood‐derived autologous therapies are safe and effective treatments for skin repair and wound healing. The fibrin network formed after plasmatic activation and the autologous growth factors released when platelets degranulate constitute a real biological medicine that has been shown to promote cell recruitment, stimulate new blood vessel formation, reduce inflammation as well as protect from local infections. This perspective highlights recent basic and clinical results published on blood‐derived autologous therapies in the field of regenerative dermatology and discusses potential challenges and future prospects.

Plasma Rich in Growth Factors Inhibits Ultraviolet B Induced Photoageing of the Skin in Human Dermal Fibroblast Culture

BACKGROUND Ultraviolet irradiation is able to deeply penetrate into the dermis and alter fibroblast structure and function, leading to a degradation of the dermal extracellular matrix. OBJECTIVES The regenerative effect of plasma rich in growth factors (PRGF) on skin ageing was investigated using UVB photo-stressed human dermal fibroblasts as an in vitro culture model. METHOD PRGF was assessed over the main indicative features of ultraviolet B irradiation, including ROS formation, cell viability and death detection, apoptosis/ necrosis analysis and biosynthetic activity measurement. Four different UV irradiation protocols were tested in order to analyze the beneficial effects of PRGF. RESULTS Ultraviolet irradiation exhibited a dose dependent cytotoxicity and dose of 400mJ/cm2 was selected for subsequent experiments. PRGF increased the cell viability and decreased the cell death comparing to the non-treated group. The apoptosis and necrosis were significantly lower in PRGF treated fibroblasts. ROS production after UV irradiation was significantly reduced in the presence of PRGF. Procollagen type I, hyaluronic acid and TIMP-1 levels were higher in the when treated with PRGF. CONCLUSION This preliminary in vitro study suggests that PRGF is able to prevent UVB derived photooxidative stress and to diminish the cell damage caused by ultraviolet irradiation.

A novel personalized 3D injectable protein scaffold for regenerative medicine

Biomaterials should be designed to closely resemble the characteristics and functions of the native extracellular matrix to provide mechanical support and signals to direct biological events. Here we have developed a novel injectable plasma rich in growth factors (PRGF-Endoret)-based formulation that combines a thermal-denaturation step of plasma with an autologous fibrin crosslinking. Rheological and mechanical properties were evaluated. Additionally, the microstructure and biological capacity of the biomaterial was also characterized. This novel formulation exhibited ideal mechanical properties and a gel-like behavior with the ability to progressively release its growth factor load over time. The results also suggested that the novel injectable formulation is non-cytotoxic, biocompatible and suitable for cell ingrowth as it is deduced from the fibroblast proliferation within the scaffold. Finally, stimulation of both cell proliferation and matrix proteins synthesis demonstrated the regenerative potential of this autologous protein based injectable scaffold.Graphical abstract

The effect of plasma rich in growth factors combined with follicular unit extraction surgery for the treatment of hair loss: A pilot study

Hair transplant surgery using follicular unit extraction technique (FUE) is a common surgical procedure for the treatment of severe hair loss. Blood‐derived autologous growth factors have also proved to promote hair regeneration in patients with different types of alopecia.

An autologous protein gel for soft tissue augmentation: in vitro characterization and clinical evaluation

The aging process affects all organs of the body, but the skin is the most visible indicator with a great psychosocial impact. As a consequence, many skin anti‐aging strategies have been developed to minimize, postpone, and even reverse the aging process. Growth factors have been widely studied in skin wound healing as these polypeptides play an essential role in the complex process of tissue regeneration. Recently, a novel 3D injectable gel based on the autologous technology platelet rich in growth factor has been described.

Platelet rich plasma for the management of hair loss: Better alone or in combination?

Platelet‐rich plasma (PRP) and autologous protein‐based treatments have recently emerged as a potential therapeutic approach for hair loss‐related disorders including androgenetic alopecia and alopecia areata. The safety and efficacy of repeated intradermal injections of PRP has proved to promote hair growth in a number of randomized clinical trials. Biologically active proteins and cytokines released upon platelet activation have shown to induce folliculogenesis and activate the anagen growing phase of dormant bulbs. Interestingly, further studies have revealed that combining PRP with other hair loss‐related products may enhance the final performance of the treatment. These synergistic approaches include Food and Drug Administration (FDA) approved drugs such as finasteride or minoxidil, bioactive macromolecules and cell‐based therapies. Here, recent research involving alone or combined therapy with platelet‐rich plasma for the management of hair loss‐related disorders are outlined and future prospects are discussed.